Analysis of killing of growing cells and dormant and germinated spores of Bacillus species by black silicon nanopillars

Ghosh, Sumana; Niu, Shanyuan; Yankova, Maya; Mecklenburg, Matthew; King, Stephen M.; Ravichandran, Jayakanth; Kalia, Rajiv K.; Nakano, Aiichiro; Vashishta, Priya; Setlow, Peter

doi:10.1038/s41598-017-18125-z

Cited by 21 publications

(15 citation statements)

References 43 publications

(78 reference statements)

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“…5b), stained red with the BacLight dye mixture, while live dormant spores do not stain well with either of the two dyes in this mixture ( Fig. 5a) (Setlow et al 2016;Ghosh et al 2017;Li et al 2017). As expected, there were significant numbers of red CTABtreated spores ( Fig.…”

Section: Baclight Staining Of Spores Ae Ctab Treatmentsupporting

confidence: 58%

“…Spores of different Bacillus strains and species incubated with CTAB or dodecylamine at various temperatures were washed with Tween-20. These spores, as well as untreated sporesAEsteam autoclave treatment for 20 min, were stained without fixation with the BacLight viability stain (ThermoFisher Scientific), and photographed by differential interference contrast (DIC) microscopy and fluorescence microscopy (Ghosh et al 2017). Phase contrast microscopy of untreated, germinated and CTAB-treated spores was carried out under 1009 magnification.…”

Section: Viability On Plates and Microscopy Of Ctab-treated Sporesmentioning

confidence: 99%

“…The BacLight Bacterial Viability stain mixture of nucleic acid dyes has also been used to assess Bacillus spore killing (Ghosh et al 2017). Thus, we also used this method to examine whether CTAB-treated spores were dead, especially as we ultimately wanted to test spores that lacked CaDPA, GRs or CLEs, and determining the viability of these mutant spores by plate counts can be difficult Paidhungat et al , 2001.…”

Section: Baclight Staining Of Spores Ae Ctab Treatmentmentioning

confidence: 99%

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Killing of spores ofBacillusspecies by cetyltrimethylammonium bromide

et al. 2019

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Aims To investigate effects of the cationic surfactant cetyltrimethylammonium bromide (CTAB), a disinfectant, on spores of Bacillus species. Methods and Results The ability of CTAB to trigger release of Bacillus spores’ large depot of dipicolinic acid (DPA) in a 1 : 1 chelate with Ca2+ (CaDPA), and to kill spores was investigated. CTAB‐triggered CaDPA release from spores of Bacillus subtilis, Bacillus cereus and Bacillus megaterium, but was not followed by completion of germination. CaDPA release triggered by CTAB increased at higher temperatures, and was optimal for B. subtilis spores at pH 9·4 and 30 μg ml−1 CTAB. CTAB also killed Bacillus spores as shown by plate counts and vital staining of treated dormant spores, and after their germination. However, B. cereus and B. megaterium spores were more CTAB‐sensitive than were B. subtilis spores. CaDPA release from and killing of CTAB‐treated spores of isogenic B. subtilis mutants lacking germination proteins was also examined, and compared with effects of the well‐known germinant dodecylamine on spores, to determine how CTAB exerts its effects on spores. Conclusions The results of this investigation showed that CTAB kills spores of three Bacillus species, perhaps by damaging the spore inner membrane, although it is also possible that some killing by this agent follows its triggering of spore germination. Significance and Impact of the Study The results of this work indicate that CTAB is also a disinfectant, but also a sporicide, and may be a useful adjunct in spore decontamination, especially at higher temperatures.

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Section: Baclight Staining Of Spores Ae Ctab Treatmentsupporting

confidence: 58%

Section: Viability On Plates and Microscopy Of Ctab-treated Sporesmentioning

confidence: 99%

Section: Baclight Staining Of Spores Ae Ctab Treatmentmentioning

confidence: 99%

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Killing of spores ofBacillusspecies by cetyltrimethylammonium bromide

et al. 2019

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“…Notwithstanding the disagreements, most researchers agree that mechanical deformation in general and the rupture of cellular components such as cell wall due to large deformations and the penetration of high aspect ratio nano-features play a key role in this regard (Fig. 3) [36,43,[53][54][55][56]. The optimum shape and dimensions of nanopatterns for maximizing the bactericidal behavior are, nevertheless, unknown (Fig.…”

Section: Bactericidal Mechanisms Of Nanopatternsmentioning

confidence: 99%

“…Subsequently, the feature size modulates the interaction of nanopatterns with cells. Not only could surface nanotopography determine stem cell fate, many studies have shown that high aspect ratio nanopatterns are capable of killing bacteria [35,36] and preventing biofilm formation [37].…”

Section: Introductionmentioning

confidence: 99%

Bactericidal effects of nanopatterns: A systematic review

et al. 2019

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We systematically reviewed the currently available evidence on how the design parameters of surface nanopatterns (e.g. height, diameter, and interspacing) relate to their bactericidal behavior. The systematic search of the literature resulted in 46 studies that satisfied the inclusion criteria of examining the bactericidal behavior of nanopatterns with known design parameters in absence of antibacterial agents. Twelve of the included studies also assessed the cytocompatibility of the nanopatterns. Natural and synthetic nanopatterns with a wide range of design parameters were reported in the included studies to exhibit bactericidal behavior. However, most design parameters were in the following ranges: heights of 100-1000 nm, diameters of 10-300 nm, and interspacings of <500 nm. The most commonly used type of nanopatterns were nanopillars, which could kill bacteria in the following range of design parameters: heights of 100-900 nm, diameters of 20-207 nm, and interspacings of 9-380 nm. The vast majority of the cytocompatibility studies (11 out of 12) showed no adverse effects of bactericidal nanopatterns with the only exception being nanopatterns with extremely high aspect ratios. The paper concludes with a discussion on the evidence available in the literature regarding the killing mechanisms of nanopatterns and the effects of other parameters including surface affinity of bacteria, cell size, and extracellular polymeric substance (EPS) on the killing efficiency. Statement of significance The use of nanopatterns to kill bacteria without the need for antibiotics represents a rapidly growing area of research. However, the optimum design parameters to maximize the bactericidal behavior of such physical features need to be fully identified. The present manuscript provides a systematic review of the bactericidal nanopatterned surfaces. Identifying the effective range of dimensions in terms of height, diameter, and interspacings, as well as covering their impact on mammalian cells, has enabled a comprehensive discussion including the bactericidal mechanisms and the factors controlling the bactericidal efficiency. Overall, this review helps the readers have a better understanding of the state-of-the-art in the design of bactericidal nanopatterns, serving as a design guideline and contributing to the design of future experimental studies.

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Nature Helps: Toward Bioinspired Bactericidal Nanopatterns

Ganjian

Modaresifar

Ligeon

et al. 2019

Adv Materials Inter

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Development of synthetic bactericidal surfaces is a drug‐free route to the prevention of implant‐associated infections. Surface nanotopographies with specific dimensions have been shown to kill various types of bacterial strains through a mechanical mechanism, while regulating stem cell differentiation and tissue regeneration. The effective ranges of dimensions required to simultaneously achieve both aims are in the <200 nm range. Here, a nanoscale additive manufacturing (=3D printing) technique called electron beam induced deposition (EBID) is used to fabricate nanopillars with reproducible and precisely controlled dimensions and arrangements that are within those effective ranges (i.e. a height of 190 nm, a diameter of 80 nm, and an interspacing of 170 nm). When compared to the flat surface, the nanopatterned surfaces show a significant bactericidal activity against both Escherichia coli and Staphylococcus aureus (with respective killing efficiencies of 97 ± 1% and 36 ± 5%). Direct penetration of nanopatterns into the bacterial cell wall leads to the disruption of the cell wall and cell death. The more rigid cell wall of S. aureus is consistent with the decreased killing efficiency. These findings support the development of nanopatterns with precisely controlled dimensions that are capable of killing both Gram‐negative and Gram‐positive bacteria.

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Analysis of killing of growing cells and dormant and germinated spores of Bacillus species by black silicon nanopillars

Cited by 21 publications

References 43 publications

Killing of spores ofBacillusspecies by cetyltrimethylammonium bromide

Killing of spores ofBacillusspecies by cetyltrimethylammonium bromide

Bactericidal effects of nanopatterns: A systematic review

Nature Helps: Toward Bioinspired Bactericidal Nanopatterns

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